[The expression of human IL-37 in E.coli and preparation and characterization of mouse anti-IL-37 antibody].
Copyright policy )[The expression of human IL-37 in E.coli and preparation and characterization of mouse anti-IL-37 antibody].
To construct a prokaryotic expression system for interleukin-37 (IL-37) and prepare its polyclonal antibody.The gene encoding mature interleukin-37 (IL-37m) was amplified by PCR and subcloned into prokaryotic expression vector pET28a. Then the recombinant plasmid pET28a/IL-37m was transformed into E.coli Rosetta and expressed under IPTG induction. The recombinant IL-37m was purified through Ni²⁺;-NT agarose gel column and the purified recombinant IL-37m was used as immunogen to immunize the BALB/c mouse. The titer and specificity of the mouse anti-IL-37 antibody were analyzed by ELISA, Western blotting and immunohistochemical staining, respectively.The recombinant IL-37 was successfully expressed and purified, and the mouse anit-IL-37 antibody was successfully prepared. ELISA showed that the titer of the antiserum was 1:128 000. Western blot analysis revealed that the antibody reacted with IL-37 specifically. Immunohistochemical staining detection manifested the antibody could recognize the native IL-37.The mouse anti-IL-37 antibody with high titer and specificity was successfully prepared.
- Guangdong Medical College China (People's Republic of)
Mice, Inbred BALB C, Blotting, Western, Gene Expression, Enzyme-Linked Immunosorbent Assay, Immunohistochemistry, Antibodies, Recombinant Proteins, Mice, Antibody Specificity, Escherichia coli, Animals, Humans, Immunization, Interleukin-1
Mice, Inbred BALB C, Blotting, Western, Gene Expression, Enzyme-Linked Immunosorbent Assay, Immunohistochemistry, Antibodies, Recombinant Proteins, Mice, Antibody Specificity, Escherichia coli, Animals, Humans, Immunization, Interleukin-1
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